WOFAPS 2025 8th World Congress of Pediatric Surgery

Shahnoor Islam ¹, AKM Amirul Morshed ²

¹ Sir salimullah medical college & Mitford hospital
² Children's Cancer Center, Dr. Azmal Hospital , Mirpur, Dhaka, Bangladesh

Background

High-quality genomic DNA from liquid and tumor tissue biopsies are necessary for biomedical research in pediatric solid tumors like nephroblastoma.

Aim

This study analyzes the qualification and purity in genomic DNA derived from liquid and tumor tissue biopsies in children with nephroblastoma.

Method

Children (0 months to 18 years) with nephroblastoma admitted in the department of pediatric surgery, Dhaka Medical College Hospital from December, 2021 to December, 2023 were included in this study with approval from Ethical Review Committee of Dhaka Medical College. Blood sample in EDTA tube and tumor tissue sample in RNA Later Solution were collected during nephrectomy after neoadjuvant chemotherapy from children with nephroblastoma. The tumor tissue biopsy used for the isolation of DNA using the QIAamp Fast DNA Tissue Kit according to the manufacturer’s protocol (Qiagen). Genomic DNA was extracted from the peripheral blood samples using the QIAamp DNA Blood Mini Kit (Qiagen). The quantity of the extracted DNA was measured using the Nanodrop. The quality of the extracted DNA was analyzed using the bioanalyzer (the bioanalyzer is a chip-based automated electrophoresis solution for the analysis of the fragment size and quality of biomolecules including RNA, DNA, protein). The DNA samples were stored at -800C before using them for studies. The ratio of absorbance at 260 and 280 nm is used to assess DNA purity. The 260/230 ratio is widely used as a secondary measure of DNA purity.

Results

Nine children with nephroblastoma were enrolled in this study. Most of the patient were unilateral localized nephroblastoma. Three children were excluded from the study after DNA extraction due to a reading below 1.6 and integrity below 7.

Conclusion

We concluded that DNA quantification and qualification is necessary for Genomic DNA in biomedical research with avoidance of high costs, low confidence results and a waste of samples.